线粒体凋亡途经参与热应激后人脐静脉内皮细胞凋亡的研究  

Mitochondrial apoptosis pathway involved in apoptosis of human umbilical vein endothelial cell after heat stress

作　　者：李莉 古正涛[1] 刘志锋[2,3] 苏磊[2,3] 

机构地区：[1]南方医科大学第三附属医院重症医学科,广州510010 [2]广州军区广州总医院重症医学科,510010 [3]全军热区创伤救治与组织修复重点实验室,510010

出　　处：《中华急诊医学杂志》2014年第12期1322-1326,共5页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（81071529,81101467）;广东省自然科学基金（10151001002000001）;广东省科技计划项目（20128031800416）;军队十二五医学科研基金重点项目（BWSl2J018）

摘　　要：目的观察细胞色素C、caspase-9和caspase-3在热应激后人脐静脉内皮细胞的表达及相互关系，阐明热应激后人脐静脉内皮细胞凋亡的信号转导机制，探讨重症中暑所致血管内皮损害的发病机理。方法建立人脐静脉内皮细胞热应激模型，对照组将细胞置于标准37℃、5％CO2细胞培养箱，热应激组将细胞置于39℃、41℃、43℃细胞培养箱中进行热应激2h，热应激后继续在细胞培养箱孵育24h。电子显微镜观察人脐静脉内皮细胞（37℃、43℃）线粒体改变，应用AnnexinV-FITC／PI双染色方法检测不同温度热应激后细胞凋亡率、Western blot检测细胞色素C、caspase-9、caspase-3蛋白表达。结果电镜观察对照组（37℃）HUVEC细胞线粒体结构基本正常。热应激后（43℃）HUVEC细胞线粒体肿胀并出现结构改变；与对照组比较，39~C热应激对内皮细胞凋亡无明显影响（P〉0．05），随着热应激温度的增加人脐静脉内皮细胞凋亡明显增多（41℃、17．8％，43°C 25．6％）并且细胞色素C、caspase-9、caspase-3蛋白表达明显增加（P〈0．05）。结论线粒体通路的激活参与了热应激后人脐静脉内皮细胞凋亡的调控；血管内皮细胞凋亡可能与重症中暑的发病存在相关性。Objective To observe the expressions of cytochrome C, Caspase-9, Caspase-3 and their relationships, and investigate apoptosis signal transduction mechanism after heat stress-induction in human umbilical vein endothelial cell （HUVEC）, and explore pathogenesis of vascular endothelial damage in the wake of severe heat stroke. Methods Human umbilical vein endothelial cell heat stress model was set up. Control group were incubated at 37℃, while heat stress group of cells were incubated at 39℃, 41℃, and 43℃ for 2h, then all the cells were further incubated at 37℃ for 24 h. Mitochondria of human umbilical vein endothelial cell were examined with electron microscopy. Apoptosis was analyzed by flow cytometry using Annexin V-FITC/PI staining, and protein levels of cytochrome C, caspase-9, caspase-3 were analyzed by western blot. Results In the control group （37℃）, the structure of mitochondrial was intact in HUVEC under transmission electron microscope. In contrast, mitochondrial swelling was found in the group of 43℃ heat stress. Compared with control group, as increasing in heat stress temperature, the rates of induced apoptosis were 17.8% at 41℃ and 25. 6% at 43℃, and the levels of cytochrome C, Caspase-9, and caspase-3 were significantly increased （P 〈0. 05）. There was no obvious change at 39℃ heat stress （P 〉 0.05）. Conclus Mitochondrial apoptosis pathway is involved in apoptosis of human umbilical vein endothelial cells in the wake of heat stress. The vascular endothelial cells apoptosis may be associated with the occurrence of severe heat stroke.

关 键 词：人脐静脉内皮细胞 凋亡 线粒体 热应激 重症中暑 细胞色素C CASPASE-9 caspase-3 

分 类 号：R594.12[医药卫生—内科学]